Figure 3

Plasma haptoglobin levels, blood–brain barrier permeability and spleen weight in mice with ECM treated with artemether with or without blood transfusion. (A) Plasma haptoglobin was evaluated using ELISA (n = 8 mice per group). Mice with ECM showed a twofold increase in plasma haptoglobin compared to uninfected control (P = 0.0074). The levels of haptoglobin were not affected by either treatment at 6 h. At 24 h, levels of haptoglobin had decreased in treated animals of both groups, being significantly different from ECM mice before treatment (P = 0.0480) and showing no significant differences in relation to uninfected control mice. (B) Permeability of blood brain barrier (BBB) was quantified by Evans blue assay. Mice with ECM showed increased permeability of the BBB in relation to uninfected controls (P = 0.0005). ECM mice treated with ARM only showed further increased permeability 6 h after treatment, but this increase was prevented in mice treated with ARM + BL. Indeed, permeability was significantly higher in ARM only-treated mice compared with mice treated with ARM + BL at 6 h (P = 0.0263). At 24 h, permeability decreased in both groups and was lower than in ECM mice before treatment (ARM only: P = 0.0065; ARM + BL: P = 0.0006), but was still higher than in uninfected controls (ARM only: P = 0.0032; ARM + BL: P = 0.0371). (C) Spleen weight: mice with ECM showed splenomegaly, with spleen weight increased more than threefold in relation to uninfected controls (P < 0.0001). Treatment with ARM only did not change spleen weight within 6 or 24 h. However, treatment with ARM + BL resulted in significant decreases in spleen weight at 6 h (P = 0.0019 and P = 0.0005 in relation to ECM mice before treatment and ARM only-treated mice, respectively) and 24 h (P = 0.0003 and P < 0.0001 in relation to ECM mice before treatment and ARM only-treated mice, respectively). However, spleen weight of ARM + BL-treated mice remained higher than uninfected controls 6 and 24 h (P < 0.0001) after treatment. Data are shown as mean ± standard deviation and Student t-test was performed for statistical analyses comparing two groups.